The project is aimed at the development of a cost effective, serum-based multiplex assay for early detection of clinically significant forms of prostate cancer. The assay utilizes metal-enhanced flourescence to provide an ultra-sensitive immunoassay for three cancer markers: alpha-methylacyl coenzyme A racemase (AMACR), annexin A3 and zinc alpha-glycoprotein (ZAG). The fluorescence enhancement is due to the plasmonic effects of a silver nanolayer in the polymer coating. The assay is adapted to PharmaSeqs innovative p-Chip platform, the key element of which is the p-Chip, a unique RFID microtransponder, which is capable of transmitting its ID via radio frequency upon being activated by light. The readout is done on a custom fluidics-based analyzer for p-Chips that simultaneously determines the fluorescence intensity and the p-Chip ID. The following tasks will be performed: 1) development of the multiplex assay involving the three markers, 2) incorporation of the silver nanolayer into the polymer coating the p-Chips to maximize the fluorescence enhancement, and 3) assay characterization and validation. The assay will be then combined with the highly sensitive, but nonspecific prostate specific antigen (PSA) test to minimize the high false positive rate. This will provide the required selectivity to eliminate hundreds of thousands of needless biopsies.